Gibson Assembly®

Gibson Assembly® was developed by Dr. Daniel Gibson and his colleagues at the J. Craig Venter Institute and licensed to NEB by Synthetic Genomics, Inc. The technique allows for the successful assembly of multiple DNA fragments, regardless of fragment length or end compatibility.

Gibson Assembly efficiently joins multiple overlapping DNA fragments in a single-tube isothermal reaction (1,2). The Gibson Assembly Master Mix includes three different enzymatic activities that perform in a single buffer:

The exonuclease creates a single-stranded 3´ overhang that facilitates the annealing of fragments that share complementarity at one end.

The polymerase fills in gaps within each annealed fragment.

The DNA ligase seals nicks in the assembled DNA.

The assembled, fully-sealed construct is then transformed into NEB 5-alpha Competent E. coli. The entire protocol, from assembly to transformation, takes a little over an hour.

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Benefits of the Gibson Assembly

Increased number of successful assembly products, particularly for longer or greater number of
fragments

Flexible sequence design (scarless cloning)

No clean-up step required

Complex assembly achieved in 1 hour

DNA can be used immediately for transformation, or as template for PCR or RCA

Easily adapted for multiple DNA manipulations, including site directed mutagenesis, insertions and
deletions

References

Here is what Customers Are Saying About Gibson Assembly

“It is very convenient and fast to clone a gene of interest into a large vector, in which unique restriction
sites are difficult to find or manipulate.”
Researcher, Washington University

“The Gibson Master Mix is a superior cloning tool that inevitably saves large blocks of time, increases
work flow, eliminates false positive clones, and saves a great deal of money! Compared to traditional
cloning, the elimination of having to produce multiple intermediate clones for multiple fragment ligations
is an enormous time saver. Furthermore, the fact that it works the first time, every time saves you from
the frustrating and time consuming troubleshooting steps. I easily saved a week’s worth screening and
subcloning with this mix. My approach to cloning anything is forever changed.”
Researcher; Sanford-Burnham Medical Research Institute

Overview of the Gibson Assembly Cloning Method

Some components of this product are manufactured by New England Biolabs, Inc. under license from Synthetic Genomics, Inc.

Legal and Disclaimers

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at gbd@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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Legal and Disclaimers

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at gbd@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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